Consequences of tributyltin chloride induced stress in Leydig cells: An ex-vivo approach

• Tributyltin chloride induces oxidative damage in Leydig cells.
• Cell death is evident which consists of both necrosis and apoptosis.
• Calcium inhibitors and free radical scavengers show protective efficacy.
• Exposure to TBTC resulted in disruption of steroidogenic activity.
• Testosterone release is significantly affected following TBTC exposure.

Tributyltin (TBT), a member of the organotin family, is a known endocrine disruptor. It persists long in the environment and is widely used in various industrial applications. This study was planned to understand its toxic influence on Leydig cells isolated from 28 day old wistar rats. In-vitro exposure to TBT-Chloride (TBTC) (300–3000 nM) reduced cell viability (DNA fragmentation, nuclear condensation and MTT assay) and affected testosterone production. TBTC induced both apoptotic and necrotic cell death (Annexin V/PI binding assay). Involvement of calcium (Ca2+), redox imbalance (ROS, GSH and TBARS) and mitochondria in TBTC toxicity was evaluated by using Ca2+ inhibitors (BAPTA-AM, EGTA, Ruthenium Red), free radical scavengers (NAC, C-Phycocyanin) and mitochondrial permeability transition pore inhibitor (Cyclosporine A). Protein expression analysis of phosphorylated MAPKinases (ERK1/2, JNK1/2, & p38), steroidogenic proteins (3β-HSD, StAR & TSPO) and apoptotic proteins (Bax, Bcl2) illustrates the cytotoxic and anti-steroidogenic activity of TBTC.